This application note monitors the existence of an intermediate state by probing the refolding of denatured cytochrome c using a JASCO CD spectrometer and the SFS-492 Stopped-Flow system.
IntroductionCD spectra provide information on the secondary structure of proteins and the environment ofaromatic side chains. Therefore, CD measurement using a stopped-flow system is considered as one ofthe best methods for analyzing the unfolding and refolding of proteins.The existence of an intermediate between denaturated state and natural state during the refolding ofproteins has been reported. The CD stopped-flow method is used for examining this refolding process. Inthis report the refolding process of cytochrome c (cyt c) measured using a SFS-492 stopped-flow systemwill be explained.Keywords: Stopped-flow, Circular Dichroism, RefoldingSample PreparationAqueous solution of Cytochrome c denaturated by guanidine hydrochloride (GuHCl) was diluted with0.1 M acetic acid buffer solution (1:9). The refolding process was observed at 222 nm for the secondary
UNC-49是一种氧化还原敏感的GABAA受体,非自主调节线粒体未折叠蛋白反应细胞UNC-49 is a redox-sensitive GABAA receptor that regulates the mitochondrial unfolded protein response cell nonautonomously